CN105661016A - Preparation method of bombus terrestris feed - Google Patents

Abstract

The invention discloses a preparation method of bombus terrestris feed. The preparation method comprises selection of strains, preparation of a yeast culture medium, preparation of a pediococcus acidilactici culture medium, preparation of a basal culture medium, preparation of a fermentation culture medium and seed culture, the fermentation culture medium is inoculated with yeast strains, culture with a fungus incubator at 28 DEG C is performed, the fermentation culture medium is heated to 50 DEG C, heat preservation is performed, and the temperature is decreased to 35 DEG C; pediococcus acidilactici strains are added, culture is performed at 35 DEG C for 1 day after even mixing, and culture at 35 DEG C is continued; feed is prepared in a sterile manner, 1/3 of the cost is reduced, worker bees with the survival time being 10-12 weeks account for 55%, the activity and disease resistance of the bombus terrestris are high, the disease resistance of the bombus terrestris is improved, and the activeness of the bombus terrestris is high; the bombus terrestris gets in and out of honeycombs 5.4 times on average every hour and is 8% higher than those independently collecting strawberry pollen and 20% higher than those independently collecting pollen and directly fed with pollen in the honeycombs in an assisted manner, the outgoing time is 4.8 hours on average every day for each bombus terrestris, and the reproductive capability of the bombus terrestris is 60% higher than that of those only collecting strawberry pollen only and 14.3% higher than those independently collecting pollen and directly fed with pollen in the honeycombs in an assisted manner.

Description

A kind of preparation method of bumblebee feedstuff

Invention field

The present invention relates to a kind of bumblebee feedstuff, specifically, relate to bumblebee (Bombusterresris) preparation method of feedstuff, belong to biological technical field.

Background technology

Bombus Apoidea insecticide, bombus, it it is a polyphagous social insect of class, there is the illumination strong, resistance to of collection power, phototaxis poor, to the features such as low temperature adaptability is strong and advantage, remain able to pollination under severe conditions, because bumblebee does not have the sensitive like that information exchanging system of Apis, wholwe-hearted crop in room temperature can gather pollination, thus becoming in greenhouse than Apis more preferably Pollinating Insect. Wan Jun etc. have studied the different pollination processing mode impact on tomato yield and quality, it was shown that bumblebee pollination processes mean fruit weight and adds 26.84% and 29.27% respectively with HORMONE TREATMENT, spontaneous pollination ratio. raising along with expanding economy and people's living standard, to anti-season vegetable and the demand of melon and fruit increasing, promote the expansion year by year of greenhouse gardening area, difference and canopy film and body of wall etc. due to planting season, indoor long-term cropping spontaneous pollination beyond affordability, traditional agriculture is pollinated frequently with artificial, mechanical vibration and hormone etc., labor intensity is big, cost and malformed fruit are high, have impact on the yield of fruit and quality, and utilize bumblebee to have developed into, for indoor crops pollination, the supplementary measures that a modern agriculture is indispensable. some agricultural developed countries of west are from beginning to research bumblebee artificial propagation techniques at the beginning of last century, and are achieved that the factorial praluction of bumblebee the eighties, it is provided that indoor agriculture pollination application. there are many large-scale scale greenhouses, especially winter in China, lacks spontaneous pollination insecticide in greenhouse, so utilizing bumblebee pollination can significantly improve yield and the quality of vegetable crop, and saves the use of labour force and medicine. owing to China's bumblebee Cultivating techniques starting is late, still there is no ripe effective bumblebee Cultivating techniques at present, only has a few countries in the world, such as Britain, Holland etc. has grasped the artificial breeding technology of bumblebee, therefore become some vegetables and fruits from external import high-quality bumblebee and plant the urgent needs of enterprise, but, difference due to home and abroad environment and cultivation of crops and pollen raw material, if entering the territory, bumblebee is fed bad on feed formula, easy feed for nursing cost is high, resistance against diseases is weak, efficiency of feed utilization is low, worker bee life cycle is short, queen to lay eggs is few, bumblebee gathers the shortcomings such as pollen adynamia, indirectly draw high the cost of bumblebee pollination, reduce bumblebee pollination efficiency, have impact on the yield of pollination crop.

Summary of the invention

The preparation method that the technical problem to be solved in the present invention is to provide a kind of bumblebee feedstuff, adopts this method gained Feed Manufacturing bumblebee, it is achieved that following purpose:

1, cost is low.

2, worker bee survival period is long.

3, queen to lay eggs is stable, and reproductive capacity is strong.

4, bumblebee liveness is high.

5, the nest time is gone out long.

6, pollination rate is high.

7, decrease artificial pollination cost, improve the yield of crop.

For solve problem above, the present invention by the following technical solutions: the preparation method of a kind of bumblebee feedstuff, it is characterised in that comprise the following steps:

A, strain are chosen: yeast and pediococcus acidilactici, 4 DEG C of inclined-plane Refrigerator stores;

B, microzyme culture medium: potato decortication stripping and slicing, every 200g adds 500mL water, boils 30min, double gauze filters to beaker, adds 20g sucrose and 5g Pollen Maydis, adds water to 1L, every 100mL solution is distributed into 250mL triangular flask, 121 DEG C of autoclaving 20min;

C, pediococcus acidilactici culture medium: take soy peptone 5.0g, Carnis Bovis seu Bubali cream powder 5.0g, yeast extract 5.0g, glucose 20g and lactose 20g, add water to 1L, and every 100mL solution is distributed into 250mL triangular flask, 121 DEG C of autoclaving 20min.

2, the preparation method of a kind of bumblebee feedstuff as claimed in claim 1, it is characterised in that further comprising the steps of:

D, basal medium: take glucose 35g, fructose 35g, sucrose 10g, Semen Glycines powder 100g, astragalus membranaceus powder 5g, Fructrs Jujubae Powder 10g, Rhizoma Dioscoreae powder 5g, Mel 20g and water 200mL, stir, load 500mL beaker, 121 DEG C of autoclaving 30min.

3, the preparation method of a kind of bumblebee feedstuff as claimed in claim 1, it is characterised in that further comprising the steps of:

E, fermentation medium: take Flos Nelumbinis pollen 150g, Flos Camelliae Japonicae powder 150g, sterilize 30min under 0.8ppm ozone/cubic meter, mixes with the basal medium cooling to 60 DEG C, controls mixture moisture 30% with sterilized water, after insulation 30min, it is cooled to room temperature and puts the preservation of 4 DEG C of refrigerator.

4, the preparation method of a kind of bumblebee feedstuff as claimed in claim 1, it is characterised in that further comprising the steps of:

F, seed culture 1: the barms taking 4 DEG C of preservations of refrigerator is inoculated into microzyme culture medium at super-clean bench, 28 DEG C, 160 revs/min of shaking tables of rotating speed are cultivated 3 days; Obtain yeast seeds;

Seed culture 2: the lactic acid bacteria taking 4 DEG C of preservations of refrigerator is inoculated into one bottle of pediococcus acidilactici culture medium at super-clean bench, 37 DEG C, 140 revs/min of shaking tables of rotating speed are cultivated 2 days, it is thus achieved that pediococcus acidilactici strain.

5, the preparation method of a kind of bumblebee feedstuff as claimed in claim 1, it is characterised in that further comprising the steps of:

G, cultured for step f yeast seeds takes 20mL being inoculated into fermentation medium, aseptic stir, 28 DEG C of fungal culture casees of temperature are cultivated 2 days.

6, the preparation method of a kind of bumblebee feedstuff as claimed in claim 1, it is characterised in that further comprising the steps of:

H, by step g cultivate after fermentation medium be warmed up to 50 DEG C be incubated 3 hours, reduce the temperature to 35 DEG C;

Add step f gained pediococcus acidilactici strain 20mL again, stir latter 35 DEG C and cultivate bottle sealing after 1 day, continues 35 DEG C of cultivations 2 days;

I, step h gains are warmed up to 55 DEG C keep 3 hours;

J, step i gains are cooled to room temperature after, sterile working divides makes column feedstuff, 4 DEG C of Refrigerator stores.

The fermentation of lactic acid bacteria, it is possible to significantly inhibit listerisa monocytogenes in mjme (see patent 2015102547559), bumblebee is mainly with sucrose solution and pollen for food, if plant pollen is less in chamber planting, it is necessary to add feed supplementation to it.In the life cycle general 4-12 week of bumblebee worker bee, length depends on the quality and quantity of pollen.

The present invention adopts above technical scheme, has the advantage that

1, cost is low, reduces by 1/3.

2, worker bee survival period is long, and life cycle 4-6 week accounts for 21%, and 7-9 week accounts for 24%, and 10-12 week accounts for 55%.

3, the activity of bumblebee, premunition are strong, and bumblebee resistance against diseases improves, and bumblebee liveness is high; Average 5.4 discrepancy Nidus Vespaes per hour, high by 8% than autonomous strawberrying pollen, gather pollen high by 20% at Nidus Vespae direct assisting in feeding pollen than autonomous.

4, technological operation is easy, is suitable to merchandized handling.

5, it is long to go out the nest time, average every day 4.8 hours/honeybee, than the raising 26.3% of second group of direct assisting in feeding pollen.

6, bumblebee reproductive capacity is than the improve 60% of a strawberrying pollen, gathers pollen the raising 14.3% at the direct assisting in feeding pollen of Nidus Vespae than autonomous.

7, gained bumblebee pollination of the present invention processes mean fruit weight and adds 29.69% and 33.98% respectively with HORMONE TREATMENT, spontaneous pollination ratio.

Below in conjunction with embodiment, the invention will be further described.

Detailed description of the invention

Embodiment 1, the preparation method of a kind of bumblebee feedstuff, comprise the following steps:

A, strain: yeast (edible Angel Yeast), 4 DEG C of inclined-plane Refrigerator stores; Pediococcus acidilactici (culture presevation CGMCCNo.10574), 4 DEG C of inclined-plane Refrigerator stores.

B, microzyme culture medium (seed culture medium): potato decortication stripping and slicing, every 200g adds 500mL water, boils 30min. Double gauze filters to beaker, adds 20g sucrose, 5g Pollen Maydis, adds water to 1L, and every 100mL solution is distributed into 250mL triangular flask, 121 DEG C of autoclaving 20min.

C, pediococcus acidilactici culture medium (seed culture medium): take soy peptone 5.0g, Carnis Bovis seu Bubali cream powder 5.0g, yeast extract 5.0g, glucose 20g, lactose 20g, add water to 1L, and every 100mL solution is distributed into 250mL triangular flask, 121 DEG C of autoclaving 20min.

D, basal medium: take glucose 35g, fructose 35g, sucrose 10g, Semen Glycines powder 100g, astragalus membranaceus powder 5g, Fructrs Jujubae Powder 10g, Rhizoma Dioscoreae powder 5g, Mel 20g, water 200mL, stir, load 3000mL triangular flask, 121 DEG C of autoclaving 30min.

E, fermentation medium: take Flos Nelumbinis pollen 150g, Flos Camelliae Japonicae powder 150g, sterilize 30min under 0.8ppm ozone/cubic meter, mixes with the basal medium cooling to 60 DEG C, controls mixture moisture 30% with sterilized water, after insulation 30min, it is cooled to room temperature and puts the preservation of 4 DEG C of refrigerator.

F, by the microzyme culture medium of preparation and pediococcus acidilactici seed culture medium, respectively 100mL/ bottle subpackage, 121 DEG C of autoclaving 20min.

G, seed culture 1: the barms taking 4 DEG C of preservations of refrigerator is placed into room temperature at super-clean bench, and sterile working is inoculated into microzyme culture medium, be placed into shaking table and cultivate, and temperature controls 28 DEG C, rotating speed 160 revs/min, incubation time 3 days; Obtain yeast seeds.

Seed culture 2: the pediococcus acidilactici strain taking 4 DEG C of preservations of refrigerator is placed into room temperature at super-clean bench, and sterile working is inoculated into pediococcus acidilactici culture medium, is placed into shaking table and cultivates, and temperature controls 37 DEG C, rotating speed 140 revs/min, incubation time 2 days; Obtain pediococcus acidilactici strain.

H, fermentation: cultured for step g barms takes 20mL and is inoculated into fermentation medium (in 3000mL triangular flask), aseptic stir, and wraps up triangular flasks with 6 layers of sterile gauze, and 28 DEG C of fungal culture casees are cultivated 2 days.

I, step h gains are warmed up to 50 DEG C be incubated 3 hours, it is ensured that yeast is killed, it is prevented that with lactic acid bacteria compete. Reduce the temperature to 35 DEG C and step g pediococcus acidilactici strain 20mL is accessed triangular flask, stir latter 35 DEG C and cultivate bottle sealing after 1 day, continues 35 DEG C of cultivations 2 days.

J, step i gains are warmed up to 55 DEG C keep 3 hours, kill pediococcus acidilactici.

K, step j gains are cooled to room temperature after, it is thick that sterile working is distributed into 1cm, 5 centimeter length column feedstuffs, 4 DEG C of Refrigerator stores.

Selected three groups of Nidus Vespaes (often 6 new Nidus Vespaes of group, each Nidus Vespae is all containing 80 workers bee) it is Experimental comparison in Fructus Fragariae Ananssae greenhouse, first group autonomous strawberrying pollen, second group independently gathers pollen and at Nidus Vespae direct assisting in feeding pollen (Flos Nelumbinis pollen and Flos Camelliae Japonicae powder each 50%), the 3rd group of autonomous strawberrying pollen at Nidus Vespae assisting in feeding gained pollen feedstuff of the present invention.

Experiment finds, first group of Nidus Vespae after 15 days average worker bee number reach 100/Nidus Vespae, reach 150/Nidus Vespae of high point after one month. Second group of Nidus Vespae after 15 days average worker bee number reach 150/Nidus Vespae, reach 210/Nidus Vespae of high point after one month. 3rd group of Nidus Vespae after 15 days average worker bee number reach 160/Nidus Vespae, ratio first group improves 60%, and ratio second group improves 6.7%; Reach 240/Nidus Vespae of high point after one month, improve 60% than first group, improve 14.3% than second group. After feeding feedstuff is described, it is possible to significantly improve the vitality of worker bee and the reproductive capacity of queen bee.

Experiment is it has furthermore been found that under same environmental condition, and first group and the 3rd group of worker bee go out nest and gather the pollen time and be substantially better than second group, first group of average every day 5.0 hours/honeybee, second group of average every day 3.8 hours/honeybee, the 3rd group of average every day 4.8 hours/honeybee. By analysis, it is short that second group of worker bee goes out the nest time, illustrates to add pollen directly to bumblebee in Nidus Vespae, makes bumblebee go out nest and gathers the adynamia of nature pollen, have impact on the function of bumblebee pollination on the contrary. Pollen can provide important energy and albumen for bumblebee, first group because not having adjuvant to supplement, worker bee needs must measuring of frequent operation collection enough every days, and excessive work and protein content deficiency are also the reasons that first group of worker bee life cycle is short. Bumblebee more likes pollen of searching for food compared with pollen feedstuff, but pollen feedstuff can provide required nutrient protein for bumblebee when pollen amount deficiency, the addition of some Chinese medicine costs, improves the resistance against diseases of the 3rd group of worker bee, makes the 3rd group of Nidus Vespae worker bee quantity ratio more than second group 14.3% simultaneously. Although the 3rd group to go out the nest time not as first group long, but first group of worker bee life-span is short, and bee colony sum is little, excessively adopts powder when pollen is under-supply, decreases the life-span of worker bee and suppresses queen bee yield, illustrates to be highly desirable to Nidus Vespae supplements.

Experiment finds simultaneously, when flower source is more, the bumblebee liveness (going out nest frequency) of the 3rd group of assisting in feeding pollen feedstuff apparently higher than first two groups, first group of single bumblebee average 5.0 discrepancy Nidus Vespae per hour, second group of average 4.5 discrepancy Nidus Vespae per hour, 3rd group of average per hour 5.4 discrepancy Nidus Vespaes, higher than first group by 8%, higher than second group by 20%, by analysis, the Chinese medicine ingredients added in pollen feedstuff improves activity and the resistance against diseases of bumblebee, increases its liveness.

Adding up by inquiry, first group of bumblebee worker bee life cycle 4-6 week accounts for 56%, and 7-9 week accounts for 33%, and 10-12 week accounts for 11%;Second group of bumblebee worker bee life cycle 4-6 week accounts for 38%, and 7-9 week accounts for 36%, and 10-12 week accounts for 26%; 3rd group of bumblebee worker bee life cycle 4-6 week accounts for 21%, and 7-9 week accounts for 24%, and 10-12 week accounts for 55%.

After repeating above test repeatedly, it has been found that result is consistent with above test.

Test post analysis, after pollen feedstuff adds Semen Glycines powder and other composition on the basis of pollen, other feed ingredient cost is low, and the manufacturing cost of pollen feedstuff can reduce by 1/3rd. The combined ferment of yeast and lactic acid bacteria, bumblebee premunition significantly improves.

The yeast and the pediococcus acidilactici that utilize domestication are breaking cellular wall strain, and technique is simple, it is easy to control. Yeast is edible yeast, and pediococcus acidilactici is screened from milk, and two kinds of bacterium clear background are transparent, environment, animal and human person will not be produced harm; Technique is simple, and index is clear, easy to operate.

Except as otherwise noted, the percent adopted in the present invention is mass percent.

Above-mentioned detailed description of the invention is illustrative of; it is to enable those skilled in the art to be better understood from present invention; should not be understood as limiting the scope of the invention, as long as according to technical solution of the present invention improvements introduced, each fall within protection scope of the present invention.

Claims (6)

1. the preparation method of a bumblebee feedstuff, it is characterised in that comprise the following steps:

A, strain are chosen: yeast and pediococcus acidilactici, 4 DEG C of inclined-plane Refrigerator stores;

B, microzyme culture medium: potato decortication stripping and slicing, every 200g adds 500mL water, boils 30min, and double gauze filters to beaker, adds 20g sucrose and 5g Pollen Maydis, adds water to 1L, 121 DEG C of autoclaving 20min;

C, pediococcus acidilactici culture medium: take soy peptone 5.0g, Carnis Bovis seu Bubali cream powder 5.0g, yeast extract 5.0g, glucose 20g and lactose 20g, add water to 1L, 121 DEG C of autoclaving 20min.

2. the preparation method of a kind of bumblebee feedstuff as claimed in claim 1, it is characterised in that further comprising the steps of:

D, basal medium: take glucose 35g, fructose 35g, sucrose 10g, Semen Glycines powder 100g, astragalus membranaceus powder 5g, Fructrs Jujubae Powder 10g, Rhizoma Dioscoreae powder 5g, Mel 20g and water 200mL, stir, 121 DEG C of autoclaving 30min.

3. the preparation method of a kind of bumblebee feedstuff as claimed in claim 1, it is characterised in that further comprising the steps of:

E, fermentation medium: take Flos Nelumbinis pollen 150g, Flos Camelliae Japonicae powder 150g, sterilize 30min under 0.8ppm ozone/cubic meter, mixes with the basal medium cooling to 60 DEG C, controls mixture moisture 30% with sterilized water, after insulation 30min, it is cooled to room temperature and puts the preservation of 4 DEG C of refrigerator.

4. the preparation method of a kind of bumblebee feedstuff as claimed in claim 1, it is characterised in that further comprising the steps of:

F, seed culture 1: the barms taking 4 DEG C of preservations of refrigerator is inoculated into microzyme culture medium at super-clean bench, 28 DEG C, 160 revs/min of shaking tables of rotating speed are cultivated 3 days; Obtain yeast seeds;

Seed culture 2: the lactic acid bacteria taking 4 DEG C of preservations of refrigerator is inoculated into one bottle of pediococcus acidilactici culture medium at super-clean bench, 37 DEG C, 140 revs/min of shaking tables of rotating speed are cultivated 2 days, it is thus achieved that pediococcus acidilactici strain.

5. the preparation method of a kind of bumblebee feedstuff as claimed in claim 1, it is characterised in that further comprising the steps of:

G, cultured for step f yeast seeds takes 20mL being inoculated into fermentation medium, aseptic stir, 28 DEG C of fungal culture casees of temperature are cultivated 2 days.

6. the preparation method of a kind of bumblebee feedstuff as claimed in claim 1, it is characterised in that further comprising the steps of:

H, by step g cultivate after fermentation medium be warmed up to 50 DEG C be incubated 3 hours, reduce the temperature to 35 DEG C;

Add step f gained pediococcus acidilactici strain 20mL again, stir latter 35 DEG C and cultivate and seal after 1 day, continues 35 DEG C of cultivations 2 days;

I, step h gains are warmed up to 55 DEG C keep 3 hours;

J, step i gains are cooled to room temperature after, sterile working divides makes column feedstuff, 4 DEG C of Refrigerator stores.